GC–MS analysis of oxysterols and their formation in cultivated liver cells (HepG2)

Oxysterols play a key role in many (patho)physiological processes and they are potential biomarkers for oxidative stress in several diseases. Here we developed a rapid gas chromatographic-mass spectrometry-based method for the separation and quantification of 11 biologically relevant oxysterols bearing hydroxy, epoxy, and dihydroxy groups. Efficient chromatographic separation (resolution ≥ 1.9) was achieved using a medium polarity 35%-diphenyl/65%-dimethyl polysiloxane stationary phase material (30 m × 0.25 mm inner diameter and 0.25 μm film thickness). Based on thorough analysis of the fragmentation during electron ionization we developed a strategy to deduce structural information of the oxysterols. Optimized sample preparation includes (i) extraction with a mixture of n-hexane/iso-propanol, (ii) removal of cholesterol by solid phase extraction with unmodified silica, and (iii) trimethylsilylation. The method was successfully applied on the analysis of brain samples, showing consistent results with previous studies and a good intra- and interday precision of ≤20%. Finally, we used the method for the investigation of oxysterol formation during oxidative stress in HepG2 cells. Incubation with tert-butyl hydroperoxide led to a massive increase in free radical formed oxysterols (7-keto-chol > 7β-OH-chol >> 7α-OH-chol), while 24 h incubation with the glutathione peroxidase 4 inhibitor RSL3 showed no increase in oxidative stress based on the oxysterol pattern. Overall, the new method described here enables the robust analysis of a biologically meaningful pattern of oxysterols with high sensitivity and precision allowing us to gain new insights in the biological formation and role of oxysterols.     

Intelligent gripper technology for the handling of carbon fiber material

Nowadays, the serial production of Carbon Fiber-Reinforced Polymers remains a challenge for the industry. Their production and application have been limited by the intensive manual work required to produce them and the resulting elevated manufacturing costs. Moreover, the production handling tasks are fulfilled to a limited extent by the gripping systems currently available in the market. The delicate process and specific material requirements of these polymers compromise the feasibility and use of automated gripper systems. An innovative solution for the automated material handling of carbon fiber textiles developed by the wbk Institute for Production Science in cooperation with J. Schmalz GmbH will be presented in this paper. The main focus of this study deals with measuring principles to increase energy efficiency, process reliability and adaptability of a gripping system using low pressure grippers. This study presents suitable solutions for the implementation of low pressure grippers in a production environment.     

An improved bacterial foraging algorithm for combined static/dynamic environmental economic dispatch

Economic dispatch is carried out at the energy control center to find out the optimal output of thermal generating units such that power balance criterion is met, unit operating limits are satisfied and the fuel cost is minimized. With growing environmental awareness and strict government regulations throughout the world, it has become essential to optimize not only the total fuel cost but also the harmful emissions, both, under static as well as dynamic conditions. The static environment economic dispatch finds the optimal output of generating units for a fixed load demand at a given time, while the dynamic environmental economic dispatch schedules the output of online generators with changing power demands over a certain time period (normally one day) so as to minimize these two conflicting objectives, simultaneously. In this paper, the price penalty factor approach is employed for simultaneous minimization of cost and emission. The generator ramp rate constraints, non-convex and discontinuous nature of cost function and the large number of generators in practical power plants, make this problem very difficult to solve. Here, a fuzzy ranking approach is employed to identify the solution which offers the best compromise between cost and emission objectives.     

Gender differences in mediated communication: Women connect more than do men

Past research in gender differences in the overall Internet use has been contradictory. Some asserted men used it more than women, while others asserted there were no gender difference. Both camps concluded that men and women differed in their motivation and utilization of time spent online. The purpose of the present research was to take a contemporary look at these gender differences. Using an online survey, we asked participants about their experiences with multiple forms of mediated communication: social networking sites, e-mail, video calls, instant messaging, texting, and phone calls. Our results indicated that women, compared to men, are generally more frequent mediated communication users. Compared to men, women prefer and more frequently use text messaging, social media, and online video calls. These results suggest that the nature of mediated social interaction is changing.     

Production and Characterization of Peptide Antibodies to the C-Terminal of Frameshifted Calreticulin Associated with Myeloproliferative Diseases

Myeloproliferative Neoplasms (MPNs) constitute a group of rare blood cancers that are characterized by mutations in bone marrow stem cells leading to the overproduction of erythrocytes, leukocytes, and thrombocytes. Mutations in calreticulin (CRT) genes may initiate MPNs, causing a novel variable polybasic stretch terminating in a common C-terminal sequence in the frameshifted CRT (CRTfs) proteins. Peptide antibodies to the mutated C-terminal are important reagents for research in the molecular mechanisms of MPNs and for the development of new diagnostic assays and therapies. In this study, eight peptide antibodies targeting the C-terminal of CRTfs were produced and characterised by modified enzyme-linked immunosorbent assays using resin-bound peptides. The antibodies reacted to two epitopes: CREACLQGWTE for SSI-HYB 385-01, 385-02, 385-03, 385-04, 385-07, 385-08, and 385-09 and CLQGWT for SSI-HYB 385-06. For the majority of antibodies, the residues Cys¹, Trp⁹, and Glu¹¹ were essential for reactivity. SSI-HYB 385-06, with the highest affinity, recognised recombinant CRTfs produced in yeast and the MARIMO cell line expressing CRTfs when examined in Western immunoblotting. Moreover, SSI-HYB 385-06 occasionally reacted to CRTfs from MPN patients when analysed by flow cytometry. The characterized antibodies may be used to understand the role of CRTfs in the pathogenesis of MPNs and to design and develop new diagnostic assays and therapeutic targets.     

A Content Analysis of Print News Coverage of Media Violence and Aggression Research

We conducted a content analysis of news articles (N = 540) to examine whether news coverage of media violence accurately reflects scientific knowledge about exposure to media violence and its effects on viewer aggression. The analysis revealed that over the past 30 years, news articles generally suggested that a link between media violence and aggression exists. However, the tone shifted sharply back toward a neutral conclusion since 2000. This shift may be attributable to the type of medium discussed (e.g., television vs. video games), the number of unaffiliated sources that are cited in the news article, and the sex of the journalist. Implications for how this news coverage may influence news readers are discussed.     

Phospholipid bilayer coatings for the separation of proteins in capillary electrophoresis

The double-chained, zwitterionic phospholipid 1,2-dilauroyl-sn-phosphatidylcholine (DLPC, C12) was investigated for its use as a wall coating for the prevention of protein adsorption in capillary electrophoresis. DLPC forms a semipermanent coating at the capillary wall, which allows excess phospholipid to be removed from the capillary prior to electrophoretic separation. A DLPC-coated capillary allowed for the separation of both cationic and anionic proteins with efficiencies as high as 1.4 million plates/m. Migration time reproducibility was less than 1.3% RSD from run to run and less than 4.0% RSD from day to day. Protein recovery was as high as 93%. Cationic and anionic proteins could be separated over a pH range of 3-10, all yielding good efficiencies (N up to 1 million plates/m). The chain length of the phospholipid affected the performance of the wall coating. The C10 analogue of DLPC (DDPC) did not form a coating on the capillary wall while the C14 analogue of DLPC (DMPC) formed a stable coating that prevented protein adsorption to the same extent as its C12 counterpart.     

The Periplasmic Oxidoreductase DsbA Is Required for Virulence of the Phytopathogen Dickeya solani

In bacteria, the DsbA oxidoreductase is a crucial factor responsible for the introduction of disulfide bonds to extracytoplasmic proteins, which include important virulence factors. A lack of proper disulfide bonds frequently leads to instability and/or loss of protein function; therefore, improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae family which is responsible for very high economic losses mainly in potato. In this work, we constructed a D. solani dsbA mutant and demonstrated that a lack of DsbA caused a loss of virulence. The mutant bacteria showed lower activities of secreted virulence determinants and were unable to develop disease symptoms in a potato plant. The SWATH-MS-based proteomic analysis revealed that the dsbA mutation led to multifaceted effects in the D. solani cells, including not only lower levels of secreted virulence factors, but also the induction of stress responses. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is crucial for D. solani virulence, and a lack of DsbA significantly disturbs cellular physiology.     

Effect of processing conditions on the yield and failure response of an aliphatic polyketone terpolymer

The effect of processing conditions on the yield and failure behavior of an aliphatic polyketone terpolymer was studied. Testing and characterization were performed on samples that were extruded in the form of hollow cylinders. We performed the extrusion process at different shear rates and at different cooling rates to assess the effect that process conditions had on the polymer properties. We performed biaxial testing on the samples to characterize the failure envelopes, including the ductile–brittle transition condition for each process condition. The effect of shear rate was negligible, whereas the cooling rate significantly affected the failure behavior. To explain these differences in behavior, we performed characterization via differential scanning calorimetry, wide‐angle X‐ray diffraction, attenuated total reflection IR spectroscopy, dynamic mechanical thermal analysis, scanning electron microscopy, and residual stress measurements. A broad glass transition was found for all samples at temperatures higher than previously reported for this material. Alteration of the processing conditions did not influence the crystalline phase (percentage crystallinity, crystalline orientation, crystallite size, etc.). A change in spherulitic structure was also observed with altered cooling rate and is suggested to have contributed to the change in failure behavior. Residual stresses also affected the behavior of all samples. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 318–334, 2002; DOI 10.1002/app.10334